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Mihaela Oprea, Maria Surdeanu, Daniela Badescu, Ani Ioana Cotar, Sorin Dinu, Otilia Banu, Mirela Flonta and Monica Straut

Abstract

The multicenter ENDOBACT study aimed at implementing molecular methods for identification of bacterial species encountered in infective endocarditis, and at attempting to reduce the number of cases with unknown etiology. For eighty seven cases was established a diagnosis of definite infective endocarditis. Thirty two of these cases had negative blood cultures. For nine cases out of 32, valve pieces were available and an attempt was made to identify the etiological agent by molecular techniques. Thirty seven available isolates were identified by phenotypical and molecular comparative methods: 16S rRNA (all available isolates), rpoB (staphylococci, streptococci and enterococci), sodA (streptococci and enterococci) genes sequencing. For eight isolates, the comparative results were discrepant. Species identification of one coagulase negative staphylococcal strain was assigned using molecular methods. Molecular identification methods applied here might represent an added value for clinical and conventional microbiological diagnosis of infective endocarditis in Romania.

Open access

Sorin Dinu, Grațiela Țârdei, Emanoil Ceaușu, Simin Aysel Florescu, Laurențiu Micu, Alina Monica Ecobici, Mariana Mihăilă and Gabriela Oprișan

Abstract

Background: Severe complications of chronic hepatitis C – i.e. cirrhosis and hepatocellular carcinoma – are important causes of morbidity and mortality worldwide. Despite the overwhelming rates of sustained virologic response achieved after therapy with different combinations of direct-acting antiviral drugs (DAAs), treatment failure is still recorded, and is due to the mutations harboured by hepatitis C virus (HCV) resistance associated variants (RAVs) selected during therapy. Baseline RAVs testing was found significant for guiding treatment in the cases of treatment failure and, sometimes, in naïve patients.

Methods: Romanian chronic hepatitis C patients unexposed to DAAs and infected with subtype 1b HCV were studied. Serum samples were used for Sanger population sequencing of a fragment containing NS3 viral protease, known to harbour resistance mutation against protease inhibitors (PIs).

Results: Catalytic triad and zinc-binding site in the studied sequences were conserved. Low-intermediate resistance mutations to first generation PIs were detected either alone or in conjunction with resistance substitutions associated with second generation PIs. Cross-resistance and reduced susceptibility to certain DAAs were observed.

Discussion: This study focused on HCV patients infected with subtype 1b strains, the most prevalent in Romania. The rate of RAVs found in this work is consistent with the results reported by similar studies from other countries. Noticeably, numerous polymorphisms of unknown significance to DAAs resistance, but reflecting the high genetic variability of HCV, were found in the studied sequences. Testing for RAVs can be a useful method for guiding treatment in a cost-efficient manner in developing countries where access to DAAs is limited.